Longevity Gene Therapy Is the Best Way to Defeat Aging

Схема-для-блога

Gene engineering is the most powerful existing tool for life extension. Mutations in certain genes result in up to 10-fold increase in nematode lifespan and in up to 2-fold increase in a mouse life expectancy. Gene therapy represents a unique tool to transfer achievements of gene engineering into medicine. This approach has already been proven successful for treatment of numerous diseases, in particular those of genetic and multigenic nature. More than 2000 clinical trials have been launched to date.

We propose developing a gene therapy that will radically extend lifespan. Genes that promote longevity of model animals will be used as therapeutic agents. We will manipulate not a single gene, but several aging mechanisms simultaneously. A combination of different approaches may lead to an additive or even a synergistic effect, resulting in a very long life expectancy. For this purpose, an animal will be affected by a set of genes that contribute to longevity. In addition, a gene therapy of all major age-related pathologies will be developed to improve the functioning of individual organs and tissues in old age. As a result, we will develop a comprehensive treatment that will not only dramatically extend lifespan, but will also prevent the decrepitude of the body. Experiments will be conducted in old mice. Thus, in case of success, the developed method of aging treatment can be quickly moved to clinical trials.

The goal of the project is to develop a complex gene therapy that will drastically increase mouse lifespan and prevent tissue pathology in old age, coupled with the safety assessment of the treatment.

Project description

11 genes that are most promising in terms of life extension (table 1) will be used as targets for gene therapy. We will affect both the biological aging mechanisms, common to all the cells of the organism, as well as the primary neuroendocrine center, that regulates the whole organism’s longevity – the hypothalamus. The expression increase or decrease of these genes in animal models was shown to result in boosted longevity. If the increase in expression of a particular gene is necessary for longevity, we will deliver this gene into the body. If, on the other hand, longevity depends on the inhibition of a certain gene’s expression, we will introduce a genetic construct that encodes small RNAs that inhibit the expression of the target gene. Two out of 10 genes have previously been used for gene therapy of aging: the lifespan of mice was increased by 20% (Zhang et al., 2013, Bernardes de Jesus et al., 2012). In addition, we will deliver 8 genes that prevent the individual tissue function disruption in old age. Each of these genes separately has previously been successfully used for gene therapy of one of the age-related diseases in rodent models (table 2).

Therapeutic genes will be introduced into the body using viral vectors – the most powerful method of delivering genetic constructs. This novel therapy that utilizes all the genes simultaniously will be used for radical life extension and for fighting decrepitude. Furthermore, each of the therapeutic genes will be tested individually. All the experiments will be conducted in 2-year old mice.

The experiments will be conducted in the following groups of experimental animals:

  • Simultaneous impact of 11 genes, known to extend life expectancy (table 1) and 8 genes that prevent the development of age-related diseases in various tissues (table 2)
  • Simultaneous impact of 11 genes, known to boost longevity (table 1)
  • Simultaneous impact of 8 genes that prevent the development of age-related pathologies in different tissues (table 2)
  • The impact of each of the 10 genes that extend lifespan, individually (11 groups of animals) (table 1)
  • Exposure to a combination of the 10 most effective geroprotectors
  • Old animals without impact
  • Young animals without impact

First of all, the efficiency of the delivery of therapeutic genes into the cells and duration of gene expression will be tested. If a tissue-specific therapy is needed, the specificity of the therapeutic construct delivery to the target tissue will be studied as well.

All groups of mice will be regularly tested for aging markers, and also the blood and adipose tissue transcriptome, proteome and metabolome will be analyzed. All age-related histological and physiological changes will be studied. Behavioral test will be performed to analyze cognitive ability and locomotor activity in mice. The average and maximum lifespan of mice will be determined. In addition, a detailed study of side effects will be performed. Mice will be compared with old mice of the control group as well as with young mice. 

Table 1. Target genes for life extending gene therapy.

Target gene

The impact on gene expression

Therapeutic effect
Effects on the hypothalamus
NF-кВ Expression inhibition

The inhibition of NF-KB transcription factor causes an increase in hormone production by the hypothalamus with during aging and hypothalamus rejuvenation

 

UCP2 Overexpression

Uncoupling protein 2 elevates the temperature of the hypothalamus, which is accompanied by a slight decrease in the overall body temperature and increased longevity

Systemic effect on most body cells
TERT Overexpression

The catalytic subunit of the telomerase extends the end regions of chromosomes – the telomeres, which increase the replicative potential of cells and longevity of the body

Repetitive sequences of the genome, encoding retrotransposons Expression inhibition

Inhibiting retrotransposon expression leads reduced genetic instability in old age

CRTC1 Expression inhibition

Inhibiting TOR-kinase, which promotes cell growth and proliferation, leads to increased life expectancy

FOXO3 Overexpression

A transcription factor that triggers stress response and promotes longevity

TFEB Overexpression

A transcription factor that activates autophagy and leads to longevity

ELAVL1 Overexpression

RNA – binding protein HuR stabilizes mRNA of factors regulating the cell cycle. The overexpression of HuR leads to rejuvenation of senescent cells

SIRT6 Overexpression

The overexpression of sirtuin 6 – a NAD + -dependent deacetylase, leads to an increase in life expectancy

AMPK genes Overexpression

AMPK overexpression triggers stress response and promotes longevity

Effect on senescent cells

HSV-TK

Overexpression

Herpes virus thymidine kinase promotes the transformation of a non-toxic prodrug into a toxic product. Thus, exposure to the prodrug induces death of senescent cells

 

Table 2. Target genes for gene therapy of age-related pathologies. Overexpression of these genes is necessary for the treatment of senile tissue decrepitude.

Target gene Tissue and delivery method

Therapeutic effect

Gene therapy research links
VEGF Systemic delivery into the blood

Vascular endothelial growth factor enhances angiogenesis (blood vessel formation)

Wang et al., 2004
BMP2, BMP7 Systemic delivery into the blood

Bone morphogenetic proteins enhance bone formation and the fracture healing process

Yue et al., 2005; Wang et al., 2008
IL-2 gene Systemic delivery into the blood

A cytokine that stimulates an immune response

Fayad et al., 2004
CREB Hippocampus

A transcription factor that in the hippocampus leads to the improvement of long-term memory formation

Mouravlev et al., 2006
IGF-1 Systemic delivery to the CNS

Insulin-like growth factor-1, whose delivery to the central nervous system (CNS) causes improvement of locomotor activity

Nishida et al., 2011
ecSOD Penis

Extracellular superoxide dismutase improves erectile function by reducing oxidative stress

Bivalacqua et al., 2003
GDNF Hypothalamus

Glial-derived neurotrophic factor that reduces obesity when delivered to the hypothalamus

Tumer et al., 2006
PVALB Heart A Ca2+- binding protein, that causes improvement of the hearts diastolic function Schmidt et al., 2005

Project authors: Anastasia Shubina, Mikhail Batin, Maria Konovalenko and Alexey Moskalev.

Literature

  1. Bernardes de Jesus B., Vera E., Schneeberger K., Tejera A.M., Ayuso E., Bosch F., Blasco M.A. Telomerase gene therapy in adult and old mice delays aging and increases longevity without increasing cancer // EMBO Mol Med. – 2012. – .v.4(8). – P.691-704.
  2. Bivalacqua T.J., Armstrong J.S., Biggerstaff J., Abdel-Mageed A.B., Kadowitz P.J., Hellstrom W.J., Champion H.C. Gene transfer of extracellular SOD to the penis reduces O2-* and improves erectile function in aged rats // Am J Physiol Heart Circ Physiol. – 2003. – v.284(4). – H1408-21.
  3. Fayad R., Zhang H., Quinn D., Huang Y., Qiao L. Oral administration with papillomavirus pseudovirus encoding IL-2 fully restores mucosal and systemic immune responses to vaccinations in aged mice // J Immunol. – 2004. – v.173(4). –P.2692-8.
  4. Mouravlev A., Dunning J., Young D., During M.J. Somatic gene transfer of cAMP response element-binding protein attenuates memory impairment in aging rats // Proc Natl Acad Sci U S A. – 2006. – v.103(12). – P.4705-10.
  5. Nishida F., Morel G.R., Hereñú C.B., Schwerdt J.I., Goya R.G., Portiansky E.L. Restorative effect of intracerebroventricular insulin-like growth factor-I gene therapy on motor performance in aging rats // Neuroscience. – 2011. – v.177. – P.195-206.
  6. Schmidt U., Zhu X., Lebeche D., Huq F., Guerrero J.L., Hajjar R.J. In vivo gene transfer of parvalbumin improves diastolic function in aged rat hearts // Cardiovasc Res. – 2005. – v.66(2). – P.318-23.
  7. Tümer N., Scarpace P.J., Dogan M.D., Broxson C.S., Matheny M., Yurek D.M., Peden C.S., Burger C., Muzyczka N., Mandel R.J. Hypothalamic rAAV-mediated GDNF gene delivery ameliorates age-related obesity // Neurobiol Aging. – 2006. – v.27(3). – P.459-70.
  8. Wang H., Keiser J.A., Olszewski B., Rosebury W., Robertson A., Kovesdi I., Gordon D. Delayed angiogenesis in aging rats and therapeutic effect of adenoviral gene transfer of VEGF // Int J Mol Med. – 2004. – v.13(4). – P.581-7.
  9. Wang Q-L., Han Q-L., Kang J., Gou S.-H., Wang L.-Zh. Polyethylenimine-mediated BMP-7 gene transfection promotes fracture healing in elderly rats // Academic Journal of Second Military Medical University. – 2008. – v.28(5). – P.514-518.
  10. Yue B., Lu B., Dai K.R., Zhang X.L., Yu C.F., Lou J.R., Tang T.T. BMP2 gene therapy on the repair of bone defects of aged rats // Calcif Tissue Int. – 2005. – v.77(6). – P.395-403.
  11. Zhang G., Li J., Purkayastha S., Tang Y., Zhang H., Yin Y., Li B., Liu G., Cai D. Hypothalamic programming of systemic ageing involving IKK-β, NF-κB and GnRH // Nature. – 2013. – v.497(7448). – P.211-6.

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Mitochondria and its role in aging

Mitochondria

 

Today was an amazing lecture by Dr. David Lee about mitochondria and its role in aging. Dr. Lee started with an overview of what mitochondria is and what it does. You may have heard that the origin of mitochondria is bacteria that was engulfed by the cell early in the course of evolution. There are several things that back this «endosymbioic» theory:

  1. Mitochondria self-replicates
  2. It has two membranes
  3. It has its own independent DNA
  4. The ribosomes are similar to those of bacteria
  5. The sizes are very alike
  6. Mitochondrial DNA shares similar to bacterial structural motifs
  7. The inner mitochondria membrane has a more bacterial-like lipid composition

Mitochondria vary from 0.5 to 10 micrometers in size. Their outer membrane is freely permeable, it let’s in and out proteins less than 5000 daltons. The inner membrane, however is tightly regulated, nothing gets in or out without the special transport. Inner membrane forms cristae that curve inside to maximize the surface for energy production.

Interestingly, there are no mitochondria in erythrocytes, however liver and muscle cells can have thousands. Mitochondria are very dynamic. They fuse and divide rapidly, however it is not known why.

Mitochondria move around to where energy is needed whithin a cell. They are transferred by kinesin-1 and dynein along the microtubules. Mitochondria are abundant close to the endoplasmatic reticulum.

Mitochondria are strictly inherited from the mother. One of the most interesting questions in biology is how the oocyte erases age-related changes from the old mitochondria during the formation of the eggs in the embryo. This rejuvenation process may hold the key to better understanding how aging can be tackled.

A mitochondrion has its one genome, which is formed by two concentric cirles – the heavy and light strains. There are 37 genes: 13 proteins, 2 ribosomal RNAs, 22 transport RNAs. 90% of the DNA is coding, with no introns and only one non-coding region, called D-loop. The genes are polycistronic, which means that one RNA molecule after transcription has several genes that are translated. All 13 proteins are from the electron transport chain, which is the energy production “factory”.

Quite amazingly, the ancestry bacteria had ~400 genes that were transferred to the nucleus, and only 13 stayed. Nobody know why these 13 and why keep its own machinery in the first place, when about 1,100 proteins come from the nucleus into the mitochondria. Moreover, pretty much all of the 13 mitochondria genes are present in the nuclear genome. They are called NUMT – nuclear mitochondria DNA. They are different than the original sequences – fragmented and scattered.

Mitochondria even have their own genetic code, which is different to what the nucleus uses. For example, AGA and AGG sequences of nucleotides in regular DNA “mean” arginine, but in the mitochondria, they “mean” stop-codone. Also, mitochondria has quite complicated transcriptome, there are small regulatory RNAs that act like siRNAs, for example, miR-1 was recently found, a nuclear RNA that regulates transcription in the mitochondria.

Mutation rate in the mitochondria is 10^4 times higher than in the nucleus and are estimated to be 6*10^-8 base pairs per year. There is a lot of debate going on about how to measure mitochondrial mutations. The methodology is much less worked through, compared to the nucleus, which makes the problem much more interesting.

The main question is whether the mitochondrial mutations come from replication infidelity, or from unrepaired damages (from ROS, for example). It can also be both, or something else, as it often happens in reality. However, this question is uite important in regard to aging, because the oxidative damage theory was one of the major ones for a while. However, now the data is accumulating that points that it may not be the dominant reason for why we age. For example, there are not so many G to T transversions that result from oxidative damage to the DNA, compared to deletions. There are also several DNA repair mechanisms: base excision repair, single-strand break repair and mismatch repair.

Then we talked about the bioenergetics – process of energy production by the mitochondria. It is called oxidative phosphorilation. Basically, the mitochondria create a difference in electrical charge between its intermembrane space and the matrix, which is inside the organelle. This difference in potential is used by complex 5 to convert ADP to ATP, cells most used fuel. More than 90% of cell’s ATP is produced in the mitochondria. They use NADH and FADH2 as proton sources. These molecules are produced inside the mitochondria in the course of nutrient processing. The nutrients are carbs, fatty acids and amino acids. Basically, mitochondria burn everything and convert to energy. I would also like to add that mitochondria are also involved in anabolic processes, meaning creating stuff – amino acids, lipids and nucleotides. Mitochondria are essenntial for proliferation.

What else do mitochondria do?

Well, they are responsible for Ca2+ ion concentration maintenance. They act like a buffer and regulate Ca2+ concentration in the cell. Normally, Ca2+ is abundant in the endoplasmatic reticulum, but it can be released in the cytoplasm, and it can go through the mitochondria. There is a uniporter that gets the ions inside the inner membrane. They are popped out by another channel, because the mitochondria have to maintain a difference in the potential.

Mitochondria regulate apoptosis. Upon breakage or leakage they release parts in the cytoplams that can be recognized as foreign material and the cell can trigger an inflammation cascade. So damaged mitochondria are very bad. There are MAVs – mitochondria anti-viral proteins that act when there’s an infection.

There is no concensus in the scientific community regarding mitochondrial DNA methylation patterns. Apparently, they are very diverse.

Mitochondria also produce heme for hemoglobin used by erythrocytes to carry oxygen.

Age-related changes in the mitochondria:

  1. More mtDNA mutations
  2. Declined respiration (ATP production)
  3. Increases ROS production
  4. Decline in fatty acid metabolism

It is not clear right now if mitochondrial DNA mutations are the source or concequence of aging.

To conclude, I’d like to say that mitochondria are an extremely interesting object for research, because their changes have a strong relationship with aging, however the details of the relationship are yet to be deciphered.

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Hollywood Must Turn Its Head to Personalized Longevity Science instead of Anti-Aging Pseudoremedies

This attention-worthy article in The Hollywood Reporter signals that Hollywood people are ready and willing to do something about their longevity. The article mentions hormone replacement therapy, different check-ups and other things available in California, however completely misses 99% of what actually can be done about aging – science. Why doesn’t the author talk about the work done at the Buck Institute for Research on Aging, USC, UCLA and Stanford University?

People are looking for a ready solution, something that they can do today, and mistakenly dismiss science completely, because they think it is too far away for being applied to them. Well, this is a wrong approach. Science can be applied to a particular person’s health. It is called personalized science. It means that we can treat a given person’s health as a scientific task. There already are several examples for personalized science in action.

Martine Rothblatt created a pharma company to invent a cure for her daughter Jenesis’s rare disease primary pulmonary hypertension, she hired the best researcher in that area back in 1996 and they created the pill that significantly improved the well-being of these patients including her daughter. This venture turned out to be quite profitable as well as being life-saving.

The other example is Michael Snyder and his recent attempt to analyze “omits” data about himself. Dr. Snyder is the Head of Genetics Department at Stanford University. He was measuring 40,000 parameters and by analyzing all this health data, his team managed to spot the onset of type 2 diabetes way earlier than he would have noticed it using conventional methods.

So, there is so much that can be done using scientific approach to health. It is not cheap, and at this point of time this kind of personalized science is for the wealthy, however the Hollywood Reporter article is exactly for this kind of crowd, it describes quite expensive health services that don’t necessarily yield results. I believe the message that science is a very powerful tool to increase longevity has to be brought to the general public, especially here in California where a great number of outstanding aging research facilities are situated.

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Biology and Biology of Aging Resources

We have prepared a list of resources that can help understand biology of aging. We tried to find easy to grasp information sources and compiled a list of lectures, audio courses, popular science books and articles on biology in general and biology of aging in particular. The selected resources probably don’t exhaust the whole picture of aging science, but they shed light on the main ideas and research directions in this area.

For those who have never studied biology systematically, we suggest to take a look at the MIT “Introduction to Biology” course.

These lectures are mainly about different aspects of molecular biology, biochemistry anad genetics. Beside that, the course provides understanding how this knowledge can be applied in real life: in gene engineering and molecular medicine.

A good online course «Useful genetics» from Coursera.org addresses genetics and its applications.

One of the lectures from this course is by the way about genetics of aging.

“Frontiers in biomedical engineering” course was presented at Yale University and is available on Youtube.

Additionally you can listen to the audio course on molecular and cell biology from UC Berkeley.

Now let’s talk about biology of aging. For starters you can read a popular science article in National Geographic titled «Longevity». It provides the most general knowledge about the genetic mechanisms that regulate aging. A very detailed description of aging on various levels (from molecular to whole organism) is provided in the books «Biology of Aging» by Roger B. McDonald and «Biology of Aging: Observations and Principles» by Robert Arking. There was a couse at MIT on biology of aging, age-related diseases and potential therapies, “The Biology of Aging: Age-Related Diseases and Interventions”, the website provides the summary. The topics that are mentioned in this course generally depict the main research avenues in biology of aging and the can be used as reference points for self studying.

Moreover, we have put together a list of lectures given by prominent scientists that provide a look at aging from different perspectives. The lecture by Vadim Gladyshev is devoted to the topic of various theories of aging – “Molecular Cause of Aging”.

Noteworthy are the lectured by the leading scientists from Stanford University, Thomas Rando and Anne Brunet  -“Longevity and Aging in Humans”

and by Janko Nikolich-Zugich from Arizona State University – “The Biology of Aging: Why Our Bodies Grow Old”

Nir Barzilai, Director of Aging Research Institute at Albert Einstein College of Medicine gave a lecture on genes, facilitating longevity in humans – “The role of protective genes in the exceptional longevity of humans” by Nir Barzilai .

Konstantin Khrapko talks about the role of mitochondia in aging – “Mitochondrial Genetics of Aging”.

Judy Campisi, professor at the Buck Institute for Research onaging, gave a talk on the relationship of cancer and cell senescence – “Cancer and aging: Rival demons”

Additionally, several lectures are about the aging of the brain – “The Aging Brain: Learning, Memory, and Wisdom,” by John Gabrieli.

and «The Aging of the Brain» by Carol A. Barnes

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Achieving Personal Immortality Roadmap

Map-Immortality_2014__ENG_31

We created the Achieving Personal Immortality Roadmap. It represents the general view on the tasks of transhumanism. It is not enough detailed, because each line requires a larger amount of texts to explain it. At some point of time we will present the plans of action on the key points and will explain how and why they need to be implemented. Nonetheless, we believe it is important to demonstrate the comprehensive view on the problem of achieving physical immortality.

The Roadmap to Personal Immortality is list of actions that one should do to live forever. The most obvious way to reach immortality is to defeat aging, to grow and replace the diseased organs with new bioengineered ones, and in the end to be scanned into a computer. This is Plan A. It is the best possible course of events. It depends on two things – your personal actions (like regular medical checkups) and collective actions like civil activism and scientific research funding. The map is showing both paths in Plan A.

However, if Plan A fails, meaning if you die before the victory over aging, there is Plan B, which is cryonics. Some simple steps can be taken now, like calling your nearest cryocompany about a contract.

Unfortunately, cryonic could also fail, and then you can move to Plan C. Of course it is much worse – less reliable and less proven. Plan C is the so called digital immortality, that means one could be returned to life based on the existing recorded information about that person. It is a not the best plan, because we are not sure how to solve the identity problem, which will arise, and also we don’t know if collected amount of information would be enough. But it is still better than nothing.

Lastly, if Plan C fails, we have Plan D. It is not a plan in fact – it is just hope or a bet that immortality already exists somehow, maybe there is quantum immortality, or maybe the future AI will bring us back to life.

All Plans demand particular actions now – we need to prepare to all of them simultaneously. All of the Plans will lead to the same result – our minds will be uploaded into a computer and will merge with AI. So these plans are in fact multilevel defense mechanisms against death structured in the most logical way.

This map is the political program of the Longevity party – this is what we are going to do. We presented the Roadmap together with Alexei Turchin near the White House as an action to increase public attention for life extension.

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Civil Activism

IMG_1597 IMG_2077 Continue reading

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Let’s Bet on Money?

labcures1

Let’s make a bet? I will propose something incredibly effective in the area of life extension and no one will be able to suggest a better strategy. Deal?

First of all, let’s think about what KPI (Key Performance Indicators) can be applied to longevity? What will tell us that we are on the right way to immortality, but not moving in the opposite direction towards the grave.

The answer here lies on the surface.

The first thing is money. The more money is invested in projects on life extesnion, the better chances we have to stay alive.

The second thing is people. There’s a nuance here. How can we count the increase in number of people? I think we need to count the most devoted supporters in the first place. There are millions of people who don’t mind living longer. Many of them run in parks or read biotechnology news, but does their activity lead to radical life extension? In my opinion, no. Moreover, having found a simple recipe like a diet, yoga or Kurzweil’s promises that everything is going to happen on its own by 2030, this kind of longevity supporters don’t do anything, because subconsciously they have already found the solution to the problem.

I propose to take only those people into account, who are involved in projects on radical human life extension. How can we identify such a person? Oh, it’s very easy. He or she talks about it explicitly and their actions are adequate, meaning they are established socially.

The third KPI is the number of events. Conferences, books, round tables, articles, social actions, 40 people going to have a beer because of the approaching singularity, youtube videos, scientific projects, movies – all of that is needed in extra large quantities.

The forth thing is the effectiveness of collaboration. This one is complicated. Perhaps the number of people involved in the same transhumanist project needs to be taken in account.

So, what strategy will be most oriented towards increasing all of the 4 indices? What can yield more money, more people, more events and more collaboration?

It is crowdfunding of scientific projects in the area of longevity. First dozens and then hundreds of crowdfunding campaigns have to be created that would raize funding for longevity gene therapy, regenerative medicine, studying molecular mechanisms of aging and their relationship to pathologies.

Yes, perhaps, we will raize not a lot of moeny in the beginning, but the projects that describe the work of the scientists, they will stay and they will continuosly owrk towards educating, promoting and attracting supporters. Moreover, they will help the universities and research institutes get government grants and venture capital.

Joint donations for longevity research will draw people closer together and will allow to constantly increase the loyal audience. There will be a chain reaction – successful crowdfunding peojects will motivate to create more projects in our field.

A set of projects on life extension will give a clear signal to the society that great things can be done to secure the main human right – the eright to live.

This is the reason why I participated in creating LabCures, because I concider this project the best strategy for the initial kick off of novel research projects including longevity.

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